Fig. 4. Effects of overlap or gap structure on the ligation by Rnl2. (A) Structural illustration of RNAs. (B) Denaturing PAGE (12%) for the ligation at 25 °C for 12 h. Lanes 1, 4 and 7: L-22a (nick), L-23a (overlap) and L-24 (2-nt gap) only, respectively. Lanes 2, 5 and 8: Rnl2 ligation of L-22a, L-23a and L-24 by Rnl2, respectively. The cyclic structures of the products in lanes 2, 5 and 8 were confirmed by their intactness against Exonuclease T in lanes 3, 6 and 9, respectively. [RNA] = 1 μM and [Rnl2] = 0.4 U μL−1 in 1× buffer.
